Effects of cashew nut shell extract and monensin on in vitro ruminal fermentation, methane production, and ruminal bacterial community.

The objective of this study was to evaluate the effects of cashew nut shell extract (CNSE) and monensin on ruminal in vitro fermentation, CH4 production, and ruminal bacterial community structure. Treatments were as follows: control (CON, basal diet without additives); 2.5 µM monensin (MON); 0.1 mg CNSE granule/g DM (CNSE100); and 0.2 mg CNSE granule/g DM (CNSE200). Each treatment was incubated with 52 mL of buffered ruminal content and 500 mg of total mixed ration for 24 h using serum vials. The experiment was performed as a complete randomized block design with 3 runs. Run was used as a blocking factor. Each treatment had 5 replicates, in which 2 were used to determine nutrient degradability, and 3 were used to determine pH, NH3-N, volatile fatty acids, lactate, total gas, CH4 production, and bacterial community composition. Treatment responses for all data, excluding bacterial abundance, were analyzed with the GLIMMIX procedure of SAS v9.4. Treatment responses for bacterial community structure were analyzed with a PERMANOVA test run with the R package vegan. Orthogonal contrasts were used to test the effects of (1) additive inclusion (ADD: CON vs. MON, CNSE100, and CNSE200); (2) additive type (MCN: MON vs. CNSE100 and CNSE200); and (3) CNSE dose (DOS: CNSE100 vs. CNSE200). We observed that pH, acetate, and acetate:propionate ratio in the CNSE100 treatment were lower compared with CNSE200, and propionate in the CNSE100 treatment was greater compared with CNSE200. Compared with MON, CNSE treatments tended to decrease total lactate concentration. Total gas production of CON was greater by 2.63% compared with all treatments, and total CH4 production was reduced by 10.64% in both CNSE treatments compared with MON. Also, compared with MON, in vitro dry matter degradabilities in CNSE treatments were lower. No effects were observed for NH3-N or in vitro neutral detergent fiber degradability. Finally, the relative abundances of Prevotella, Treponema, and Schwartzia were lower, whereas the relative abundances of Butyrivibrio and Succinivibrio were greater in all treatments compared with CON. Overall, the inclusion of CNSE decreased CH4 production compared with MON, making CNSE a possible CH4 mitigation additive in dairy cattle diets.

Production, physiological response, and calcium and magnesium balance of lactating Holstein cows fed different sources of supplemental magnesium with or without ruminal buffer.

The objective of this study was to evaluate the effects of dietary replacement of magnesium oxide (MgO) with calcium-magnesium hydroxide [CaMg(OH)2] and its interaction with ruminal buffer (sodium sesquicarbonate) supplementation on production, Ca and Mg balance, and overall physiological response of mid-lactation Holstein dairy cows. Sixty cows averaging 40.5 ± 7.0 kg of milk/d were used. Treatments were assigned following a 2 × 2 factorial arrangement: (1) MgO, (2) MgO + buffer, (3) CaMg(OH)2, or (4) CaMg(OH)2 + buffer. Diets were formulated to have 16.5% of crude protein, 1.82 Mcal/kg of net energy for lactation, 0.67% Ca, 0.39% P, and 0.25% Mg, all on a dry matter (DM) basis. Treatments were individually top dressed. Milk production, composition, and DM intake were evaluated. A subsample of 20 cows were randomly selected for the evaluation of Ca and Mg balance, blood gases, and electrolytes. Ruminal fluid was also collected for evaluation of pH and Ca and Mg solubility. Effects of Mg source, buffer, and the interaction Mg source × buffer were analyzed through orthogonal contrasts. An interaction of Mg source × buffer was found for DM intake and feed efficiency, in which cows fed CaMg(OH)2 had a similar feed efficiency regardless of ruminal buffer inclusion; however, when cows were fed MgO, the inclusion of buffer reduced feed efficiency. No effects on body weight and milk yield were observed. Buffer addition tended to increase the concentrations of fat, protein, and solids-not-fat, without affecting the yields of these milk components. Magnesium source and buffer did not affect ruminal fluid, blood, urine, or fecal pH; however, buffer supplementation increased urinary pH. Treatment with CaMg(OH)2 increased blood concentration of HCO3-, total CO2, and base excess compared with cows fed MgO. No differences were observed in the ruminal solubility of Ca and Mg or on milk or urinary Ca and Mg excretion. Greater plasma Mg concentration was observed for animals fed MgO compared with cows fed CaMg(OH)2; however, both sources were above the threshold recommended in the literature for dairy cows. Also, a reduction in fecal Mg excretion was observed in animals fed CaMg(OH)2. In summary, we provide evidence that CaMg(OH)2 could replace MgO without affecting performance, overall physiological response, or Ca and Mg balance of mid-lactating dairy Holstein cows.

Effects of choline chloride on the ruminal microbiome at 2 dietary neutral detergent fiber concentrations in continuous culture.

Our objective was to evaluate the effects of unprotected choline chloride (Cho) on the ruminal microbiome at 2 dietary neutral detergent fiber (NDF) concentrations. We hypothesized that the effects of Cho on ruminal bacterial populations would depend on NDF. Eight dual-flow continuous-culture fermentors were arranged in a duplicated 4 × 4 Latin square as a 2 × 2 factorial with the following treatments: (1) 30% NDF-control (30% NDF diet, no supplemental choline); (2) 30% NDF-Cho (30% NDF diet plus 1.9 g of choline ion per kg of dry matter); (3) 40% NDF-control (40% NDF diet, no supplemental choline); and (4) 40% NDF-Cho (40% NDF diet plus 1.9 g of choline ion per kg of dry matter). We did 4 fermentation periods of 10 d each and used the last 3 d for collection of samples of solid and liquid digesta effluents for DNA extraction. Overall, 32 solid and 32 liquid samples were analyzed by amplification of the V4 variable region of bacterial 16S rRNA. Data were analyzed with R (R Project for Statistical Computing) and SAS (SAS Institute Inc.) to determine effects of Cho, NDF, and NDF × Cho on taxa relative abundance. The correlation of propionate molar proportion with taxa relative abundance was also analyzed. At the phylum level, relative abundance of Firmicutes in the liquid fraction tended to be greater when Cho was supplemented with a 30% NDF diet. At the order level, Cho increased Coriobacteriales in solid fraction and decreased Fibrobacterales in liquid fraction. Moreover, Cho decreased abundance of Clostridiales and increased Selenomonadales in the solid fraction, only with the 30% NDF diet. For genera, lower abundance of Pseudobutyrivibrio resulted from Cho in solid and liquid fractions. Greater abundance of Succinivibrio in solid and Selenomonas and Selenomonas 1 in liquid resulted from Cho with the 30% NDF diet. Propionate molar proportion was positively correlated with relative abundance of order Selenomonadales in solid and liquid fractions, and with genus Succinivibrio in solid and genera Selenomonas and Selenomonas 1 in liquid. Our results indicate that Cho primarily decreases abundance of bacteria involved in fiber degradation and increases abundance of bacteria mainly involved in nonstructural carbohydrate degradation and synthesis of propionate, particularly when a diet with 30% NDF is provided.

Effects of replacing magnesium oxide with calcium-magnesium carbonate with or without sodium bicarbonate on ruminal fermentation and nutrient flow in vitro.

The objective of this study was to evaluate the effects of replacing magnesium oxide (MgO) with calcium-magnesium carbonate [CaMg(CO3)2] on ruminal fermentation with or without the addition of sodium bicarbonate (NaHCO3). Eight fermentors of a dual-flow continuous-culture system were distributed in a replicated (2) 4 × 4 Latin square design in a 2 × 2 factorial arrangement of treatments (magnesium sources × NaHCO3). The treatments tested were 0.21% MgO [MgO; dry matter (DM) basis; 144.8 mEq of dietary cation-anion difference (DCAD)]; 0.21% MgO + 0.50% NaHCO3 (MgO+NaHCO3; DM basis; 205.6 mEq of DCAD); 1.00% CaMg(CO3)2 [CaMg(CO3)2; DM basis; 144.8 mEq of DCAD]; and 1.00% CaMg(CO3)2 + 0.50% NaHCO3 [CaMg(CO3)2+NaHCO3; DM basis; 205.6 mEq of DCAD]. Diets were formulated to have a total of 0.28% of Mg (DM basis). The experiment consisted of 40 d, which was divided into 4 periods of 10 d each, where 7 d were used for adaptation and 3 d for sampling to determine pH, volatile fatty acids (VFA), ammonia (NH3-N), lactate, mineral solubility, N metabolism, and nutrient digestibility. The effects of Mg source [MgO vs. CaMg(CO3)2], NaHCO3 (with vs. without), and the interaction were tested with the MIXED procedure of SAS version 9.4 (SAS Institute). There was no Mg source × NaHCO3 interaction in the pH variables and mineral solubility, and Mg sources evaluated did not affect the variables related to ruminal pH and solubility of Mg. On the other hand, the inclusion of NaHCO3 increased the pH daily average, independent of Mg source, which led to a reduced time that pH was below 5.8 and decreased area under the curve. Total VFA and lactate concentration were similar among treatments regardless of NaHCO3 and Mg source; however, the molar proportion of isobutyrate and NH3-N concentration were lower in diets with CaMg(CO3)2 compared with MgO. Moreover, NaHCO3 inclusion increased NH3-N, total daily NH3-N flow, isobutyrate concentration, and acid detergent fiber digestibility. Our results showed that CaMg(CO3)2 leads to a lower NH3-N concentration and isobutyrate proportion. Therefore, because most of the tested variables were not significantly different between MgO and CaMg(CO3)2 when combined or not with NaHCO3, CaMg(CO3)2 can be a viable alternative source to replace MgO in dairy cow diets without affecting mineral solubility, ruminal pH, nutrient digestibility, total VFA, and the main ruminal VFA. Although Mg sources are known to have an alkalizing effect, NaHCO3 inclusion in diets with Mg supplementation allowed an increase in ruminal pH, as well as an increase in isobutyrate and NH3-N flow.

Shrink, weighing accuracy, and weighing precision of mineral supplement in five commercial dairies in the Western United States.

Shrink, accuracy, and precision of ingredient weighing are critical factors of efficiency in TMR-fed dairy systems. Those factors have been evaluated for major feeds; however, we are not aware of any reports for mineral supplement. Farms commonly mix mineral supplement with other low-inclusion ingredients into a premix which is used later as a single ingredient for TMR formulation. Our objectives were to evaluate shrink, weighing accuracy, and weighing precision of mineral supplement during premix formulation, and variation in concentration of minerals in mineral supplement and TMR, in five large dairies in the Western United States. We used the automated weight-tracking system at each farm to account for all the mineral supplement loaded into the mixing-wagon and collected samples of mineral supplement and TMR from time of mineral supplement delivery at the farm until 100% of it was consumed. Mean, standard deviation and coefficient of variation (CV) for each variable were calculated with SAS 9.4. Average shrink was estimated at 2.0% for mineral supplement during storage and loading, ranging from 0.37% to 3.25%. Mineral supplement weighing deviation from the targeted amount was 1.54% on average for the five dairies with a 95% CV. Mineral composition of mineral supplements averaged 11.3%, 0.27%, and 3.16% for Ca, P, and Mg, and 215, 881, and 1533 ppm for Cu, Mn, and Zn, respectively. Mineral compositions in TMR averaged 0.84%, 0.41%, and 0.37% for Ca, P, and Mg, respectively; and 15.1, 71, and 94.5 ppm for Cu, Mn, and Zn, respectively. The CV of all minerals except Ca, were larger for mineral supplement than TMR, and with the exception of P in mineral supplement, CV of trace minerals were larger than CV values for macro minerals. Our shrink estimates for mineral supplement represent an initial approximation to this issue. Results of our weighing deviation analysis suggest some room for improvement on the precision of weighing mineral supplement at the time when premix is prepared at the farm, which could improve consistency in chemical composition of the premix and consequently reduce the variation (CV values) of mineral concentrations in TMR.

Effects of supplemental source of magnesium and inclusion of buffer on ruminal microbial fermentation in continuous culture.

Magnesium oxide (MgO) is the most common supplemental source of Mg for dairy cows and a proven ruminal alkalizer when supplemented above NRC (2001) recommendations. However, overfeeding MgO may increase feeding costs, whereas the effects of alternative sources of Mg on ruminal fermentation are not well known. Moreover, it is still unclear if Mg supplementation influences the effects of bicarbonate-based buffers on ruminal fermentation. We aimed to evaluate the effect of Mg source on ruminal fermentation with diets formulated to a final concentration of 0.25% Mg, and to determine if the effect of sodium sesquicarbonate as a buffer varies with the source of Mg. We used 8 fermentors in a duplicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments, by combining 2 factors: (1) Mg source: using either MgO or an alternative source consisting of a blend of CaMg(OH)4 and CaMg(CO3)2 (BLN) and (2) sodium sesquicarbonate buffer inclusion, at 0 or 0.6% of dry matter intake. Based on preliminary tests of reactivity, we hypothesized that BLN plus buffer would allow for greater ruminal pH, acetate molar proportion, and NDF digestibility than diets with MgO or without buffer. Four 10-d periods were completed, where the last 3 d were used for pH measurements and collection of samples for volatile fatty acids (VFA), ammonia (NH3-N), Mg solubility, N metabolism, and nutrient digestibility. Effects of Mg source (source), sodium sesquicarbonate inclusion (buffer), and their interaction (source × buffer) were tested with the MIXED procedure of SAS (SAS Institute Inc.). We did not find an effect of Mg source on ruminal fermentation variables; however, concentration of soluble Mg in ruminal fluid was greater for MgO compared with BLN. On the other hand, buffer supplementation increased average ruminal pH, acetate molar proportion, and branched-chain VFA molar proportion; tended to increase NDF digestibility; and decreased both area under the curve and time below pH 6.0. An interaction of source × buffer was found for propionate, butyrate, and NH3-N, the first one decreasing and the 2 others increasing only when buffer was supplemented to the BLN diet. Our results indicate that supplementing Mg with either MgO or BLN promotes similar ruminal fermentation in diets with total concentration of 0.25% Mg. Further evaluations are needed to assess Mg availability and animal performance in dairy cows fed BLN.

Effect of increasing dietary protein with constant lysine:methionine ratio on production and omasal flow of nonammonia nitrogen in lactating dairy cows.

Eight lactating cows were fed 4 diets in which dietary crude protein (CP) was increased in steps of approximately 2 percentage units from 11 to 17% of DM by replacing high-moisture corn with soybean meal supplemented with rumen-protected Met to maintain a Lys:Met ratio of 3:1 in metabolizable protein. Trial design was a replicated 4 × 4 Latin square; experimental periods lasted 28 d, with data and sample collection being performed during wk 3 and 4 of each period. Digesta samples were collected from the rumen as well as the omasum to measure metabolite concentrations and ruminal outflow of N fractions using infusion of 15N-enriched ammonia to quantify microbial nonammonia N (NAN) and nonmicrobial NAN. Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc.). There were linear increases in the yields of milk and true protein and concentration of milk urea N, and a linear decrease in N efficiency, with increasing dietary CP. Apparent ruminal and total-tract N digestibility increased linearly with increasing dietary CP, but estimated true total-tract N digestibility was not affected. Apparent digestibility of the other macronutrients was not influenced by diet. Ruminal ammonia, total AA and peptides, and branched-chain VFA also increased linearly with dietary CP. The 15N enrichment of liquid- and particle-associated microbes linearly declined with increasing dietary CP due to decreasing 15N enrichment of the ammonia pool. Although no effect of dietary CP on nonmicrobial NAN flow was detected, total NAN flow increased linearly from 525 g/d at 11% CP to 637 g/d at 17% CP due to the linear increase in microbial NAN flow from 406 g/d at 11% CP to 482 g/d at 17% CP. Under the conditions of this study, when dietary CP was increased by adding soybean meal supplemented with rumen-protected Met, improved milk and protein yields were driven not by RUP supply but by increased ruminal outflow of microbial protein.

Effects of calcium-magnesium carbonate and calcium-magnesium hydroxide as supplemental sources of magnesium on microbial fermentation in a dual-flow continuous culture.

Supplemental sources of Mg can also aid in ruminal pH regulation due to their alkaline properties. Magnesium oxide (MgO) is the most common source of Mg for ruminants and can help controlling ruminal pH; however, the alkaline potential of other sources of Mg has not been evaluated. We aimed to evaluate the inclusion of calcium-magnesium carbonate (CaMg(CO3)2) and calcium-magnesium hydroxide (CaMg(OH)4) alone or in combination as supplemental sources of Mg in corn silage-based diets and its impact on ruminal microbial fermentation. We hypothesized that inclusion of CaMg(OH)4 would allow for ruminal fermentation conditions resulting in a greater pH compared to the inclusion of CaMg(CO3)2. Four treatments were defined by the supplemental source of Mg in the diet: 1) Control (100% MgO, plus sodium sesquicarbonate as a buffer); 2) CO3 [100% CaMg(CO3)2]; 3) OH [100% CaMg(OH)4]; and 4) CO3/OH [50% Mg from CaMg(CO3)2, 50% Mg from CaMg(OH)4]. Nutrient concentration was held constant across treatments (16% CP, 30% NDF, 1.66 Mcal NEl/kg, 0.67% Ca, and 0.21% Mg). Four fermenters were used in a 4 × 4 Latin square design with four periods of 10 d each. Samples were collected for analyses of nutrient digestibility, soluble Mg, VFA, and NH3, while pH was measured at 0, 1, 2, 4, 6, 8, and 10 h post morning feeding to estimate % time when pH was below 6 (pH-B6) and area under the pH curve for pH below 6.0 (pH-AUC). Bacteria pellets were harvested for 15N analysis and estimates of N metabolism. Treatment effects were analyzed with the mixed procedure of SAS, while effects of using either CaMg(CO3)2 or CaMg(OH)4 as Mg source in comparison to Control treatment were evaluated by orthogonal contrasts. Similar pH-related variables were observed for Control, OH, and CO3/OH treatments, which had smaller pH-AUC and pH-B6 than CO3 (P ≤ 0.01). Butyrate molar proportion was greater in Control and CO3/OH than in CO3 and OH (P = 0.04). Orthogonal contrasts showed lower flow of bacterial N (P = 0.04), lower butyrate molar proportion (P = 0.08) and greater pH-AUC (P = 0.05) for diets with CaMg(CO3)2 in comparison with the Control. Concentration of soluble Mg in ruminal fluid (P = 0.73) and nutrient digestibility (P ≥ 0.52) were similar across treatments. Under the conditions of this experiment, using CaMg(OH)4 alone or combined with CaMg(CO3)2 allowed for a less acidic ruminal fermentation pattern than a diet with only CaMg(CO3)2.

Effects of unprotected choline chloride on microbial fermentation in a dual-flow continuous culture depend on dietary neutral detergent fiber concentration.

Choline is usually supplemented as ruminally protected choline chloride to prevent its degradation in the rumen, but the effects of unprotected choline on ruminal fermentation are unclear. Some research indicates a possible role of dietary fiber on microbial degradation of choline; therefore we aimed to evaluate the effects of unprotected choline chloride on ruminal fermentation and to investigate whether those effects depend on dietary neutral detergent fiber (NDF) concentration. Our hypothesis was that dietary NDF concentration would influence choline chloride effects on microbial ruminal fermentation. We used 8 fermentors in a duplicated 4 × 4 Latin square with a 2 × 2 factorial arrangement, combining 2 factors: (1) dietary NDF concentration and (2) unprotected choline chloride supplementation. Resulting treatments are (1) 30%NDF/Ctrl [30% NDF control diet without supplemental choline (Cho)]; (2) 30%NDF/Cho [30% NDF diet plus 1.9 g of choline ion per kg of dry matter (DM)]; (3) 40%NDF/Ctrl (40% NDF control diet without supplemental choline); and (4) 40%NDF/Cho (40% NDF diet plus 1.9 g of choline ion per kg of DM). Four 10-d periods were completed, each consisting of 7 d for adaptation and 3 d for collection of samples for estimation of nutrient disappearance and daily average concentrations of volatile fatty acids and NH3-N. In addition, kinetics of pH, acetate, and propionate were evaluated at 0, 1, 2, 4, 6, and 8 h after morning feeding. On the last day of each period, bacteria pellets were harvested for 15N analysis and N metabolism. Fixed effects of dietary NDF concentration, unprotected choline chloride supplementation, and their interaction (NDF × Cho) were tested using the MIXED procedure of SAS version 9.4 (SAS Institute Inc., Cary, NC). Choline tended to increase total volatile fatty acid concentrations and decreased acetate molar proportion regardless of dietary NDF concentration, but it increased propionate molar proportion and decreased acetate to propionate ratio only with the 30% NDF diet. Supplementing choline decreased NDF disappearance regardless of dietary NDF; however, organic matter disappearance tended to be reduced only when choline was added to 40% NDF. Our data indicate that unprotected choline chloride effects on ruminal fermentation depend on dietary NDF concentration, allowing for a greater propionate synthesis without decreasing organic matter disappearance when fed with a 30% NDF diet.

Copper sulfate and sodium selenite lipid-microencapsulation modifies ruminal microbial fermentation in a dual-flow continuous-culture system.

Undesirable interactions between trace mineral elements and ruminal contents may occur during digestion when mineral salts are supplemented. Antimicrobial effects of copper sulfate (CuSO4) may affect ruminal digestibility of nutrients when fed as a source of copper (Cu), while sodium selenite (Na2SeO3) may be reduced in the rumen to less available forms of selenium (Se). Our objective was to evaluate if protection of CuSO4 and Na2SeO3 by lipid-microencapsulation would induce changes on ruminal microbial fermentation. We used 8 fermentors in a dual-flow continuous-culture system in a 4 × 4 duplicated Latin square with a 2 × 2 factorial arrangement of treatments. Factors were CuSO4 protection (unprotected and protected by lipid-microencapsulation) and Na2SeO3 protection (unprotected and protected by lipid-microencapsulation). Treatments consisted of supplementation with 15 mg/kg of Cu and 0.3 mg/kg of Se from either unprotected or protected (lipid-microencapsulated) sources, as follows: (1) Control (unprotected CuSO4 + unprotected Na2SeO3); (2) Cu-P (protected CuSO4 + unprotected Na2SeO3); (3) Se-P (unprotected CuSO4 + protected Na2SeO3); (4) (Cu+Se)-P (protected CuSO4 + protected Na2SeO3). All diets had the same nutrient composition and fermentors were fed 106 g of dry matter/d. Each experimental period was 10 d (7 d of adaptation and 3 d for sample collections). Daily pooled samples of effluents were analyzed for pH, NH3-N, nutrient digestibility, and flows (g/d) of total N, NH3-N, nonammonia N (NAN), bacterial N, dietary N, and bacterial efficiency. Kinetics of volatile fatty acids was analyzed in samples collected daily at 0, 1, 2, 4, 6, and 8 h after feeding. Main effects of Cu protection, Se protection, and their interaction were tested for all response variables. Kinetics data were analyzed as repeated measures. Protection of Cu decreased acetate molar proportion, increased butyrate proportion, and tended to decrease acetate:propionate ratio in samples of kinetics, but did not modify nutrient digestibility. Protection of Se tended to decrease NH3-N concentration, NH3-N flow, and CP digestibility; and to increase flows of nonammonia N and dietary N. Our results indicate that protection of CuSO4 may increase butyrate concentration at expenses of acetate, while protection of Na2SeO3 tended to reduce ruminal degradation of N. Further research is needed to determine the effects of lipid-microencapsulation on intestinal absorption, tissue distribution of Cu and Se, and animal performance.

Effects of replacing soybean meal with canola meal for lactating dairy cows fed 3 different ratios of alfalfa to corn silage.

Previous research has demonstrated that feeding canola meal (CM) improves milk production and N utilization by lactating dairy cows when replacing solvent-extracted soybean meal (SBM). The objective of the present study was to evaluate whether CM would improve milk yield and components and N utilization, compared with SBM, at different ratios of alfalfa silage (AS) to corn silage (CS) fed to lactating dairy cows. Twenty-four multiparous Holstein cows averaging, at the beginning of the study (mean ± SD), 2.8 ± 0.9 parity, 684 ± 56 kg of BW, 102 ± 41 DIM, and 49 ± 4 kg milk/d, and 24 primiparous cows averaging (mean ± SD) 565 ± 46 kg of BW, 123 ± 30 DIM, and 40 ± 4 kg milk/d were blocked by parity and DIM. A cyclic changeover design with 4 replications of 2 blocks of treatments of 6 cows was used in an arrangement with 4 28-d periods. Dietary treatments were arranged in a 3 × 2 factorial design of 3 proportions of AS to CS as forage source (HAS = high AS, 50% AS to 10% CS; MAS = medium AS, 30% AS to 30% CS; LAS = low AS, 10% AS to 50% CS) and 2 protein supplements (CM vs. SBM). Diets were formulated to contain [dry matter (DM) basis]: 60% forage, 8 to 15% high-moisture corn, 2 to 5% soy hulls, 1.3% mineral-vitamin premix, 16% crude protein, and 31 to 33% NDF. Data from the last 2 weeks of each period were used to compute mean milk yield and composition, and efficiencies of feed conversion, for each cow in each period. Data for the other variables were collected during the last week of each period. All data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Regardless of the forage source, replacing SBM with CM improved yields of milk, milk protein, and solids-not-fat. Moreover, milk urea nitrogen concentration and urinary excretion of total N (g/d) and urea N (% of total urinary N) decreased when CM replaced SBM. An interaction effect occurred between forage source and protein supplements for apparent total-tract digestibility, and, overall, this effect was due to small differences in ingredient and chemical compositions of the diets. In addition, these differences had a minor effect on cow performance. Yields of milk and milk components were greatest for cows fed 50% CS, intermediate for 30% CS, and lowest for 10% CS, indicating that, under the conditions of the present study, cows fed 50% CS in the diet (DM basis) had greater production compared with those fed 50% AS.

Effects of methionine plus cysteine inclusion on performance and body composition of liquid-fed crossbred calves fed a commercial milk replacer and no starter feed.

This experiment aimed to evaluate the effects of supplying 4 different inclusion levels of Met + Cys to crossbred liquid-fed calves on animal performance and body composition. Thirty-six Holstein-Gyr male calves were separated into 2 age groups: 16 calves, slaughtered at an age of 30 d, representing the physiological phase from 8 to 30 d, and 20 calves, slaughtered at an age of 60 d, representing the physiological phase from 30 to 60 d. At 8 d of age, the animals were randomly distributed among the experimental treatments: 4 Met + Cys inclusion levels (Met + Cys: 8.0, 8.7, 9.4, and 10.2 g/d), provided by an AA supplement added to 1.0 kg (as fed) of commercial milk replacer containing soy protein concentrate and wheat protein isolate reconstituted at 13.8% (dry matter basis). The diet was supplied without allowing leftovers and no starter feed was provided. The experimental diets were supplied without allowing orts, so that the dry matter, crude protein, and ether extract intakes were the same for all animals, independent of Met + Cys level. Total weight gain, average daily gain, gain composition, and body composition were evaluated for both age groups separately. Digestibility of organic matter, crude protein, and ether extract was lower for 8 to 30 d than for 30 to 60 d. The effect of Met + Cys levels on the digestibility of nutrients was not observed; there also was no significant interaction between physiological phase and Met + Cys levels. For the 8 to 30 d group, no responses in performance were observed according to the different Met + Cys levels, which indicates that 8.0 g/d of Met + Cys met the requirements for this physiological phase. The 30 to 60 d group responded positively to higher Met + Cys inclusion in the diet. In conclusion, an optimal Met + Cys dietary level to ensure best performance and protein gain ranges from 8.41 to 9.81 g/d.

Effect of replacing calcium salts of palm oil with camelina seed at 2 dietary ether extract levels on digestion, ruminal fermentation, and nutrient flow in a dual-flow continuous culture system.

Camelina is a drought- and salt-tolerant oil seed, which in total ether extract (EE) contains up to 74% polyunsaturated fatty acids. The objective of this study was to assess the effects of replacing calcium salts of palm oil (Megalac, Church & Dwight Co. Inc., Princeton, NJ) with camelina seed (CS) on ruminal fermentation, digestion, and flows of fatty acids (FA) and AA in a dual-flow continuous culture system when supplemented at 5 or 8% dietary EE. Diets were randomly assigned to 8 fermentors in a 2 × 2 factorial arrangement of treatments in a replicated 4 × 4 Latin square design, with four 10-d experimental periods consisting of 7 d for diet adaptation and 3 d for sample collection. Treatments were (1) calcium salts of palm oil supplementation at 5% EE (MEG5); (2) calcium salts of palm oil supplementation at 8% EE (MEG8); (3) 7.7% CS supplementation at 5% EE (CS5); and (4) 17.7% CS supplementation at 8% EE (CS8). Diets contained 55% orchardgrass hay, and fermentors were fed 72 g of dry matter/d. On d 8, 9, and 10 of each period, digesta effluent samples were taken for ruminal NH3, volatile fatty acids, nitrogen metabolism analysis, and long-chain FA and AA flows. Statistical analysis was performed using the MIXED procedure (SAS Institute Inc., Cary, NC). We detected an interaction between FA source and dietary EE level for acetate, where MEG8 had the greatest molar proportion of acetate. Molar proportions of propionate were greater and total volatile fatty acids were lower on CS diets. Supplementation of CS decreased overall ruminal nutrient true digestibility, but dietary EE level did not affect it. Diets containing CS had greater biohydrogenation of 18:2 and 18:3; however, biohydrogenation of 18:1 was greater in MEG diets. Additionally, CS diets had greater ruminal concentrations of trans-10/11 18:1 and cis-9,trans-11 conjugated linoleic acid. Dietary EE level at 8% negatively affected flows of NH3-N (g/d), nonammonia N, and bacterial N as well as the overall AA outflow. However, treatments had minor effects on individual ruminal AA digestibility. The shift from acetate to propionate observed on diets containing CS may be advantageous from an energetic standpoint. Moreover, CS diets had greater ruminal outflow of trans-10/11 18:1 and cis-9,trans-11 conjugated linoleic acid than MEG diets, suggesting a better FA profile available for postruminal absorption. However, dietary EE at 8% was deleterious to overall N metabolism and AA outflow, indicating that CS can be fed at 5% EE without compromising N metabolism.

Lactation response to soybean meal and rumen-protected methionine supplementation of corn silage-based diets.

Corn silage, an important forage fed to dairy cows in the United States, is energy rich but protein poor. The objectives of this experiment were to investigate the effects on production of milk and milk components of feeding corn silage-based diets with 4 levels of dietary crude protein (CP) plus rumen-protected methionine (RPM). Thirty-six cows were blocked by days in milk into 9 squares and randomly assigned to 9 balanced 4 × 4 Latin squares with four 4-wk periods. All diets were formulated to contain, as a percent of dry matter (DM), 50% corn silage, 10% alfalfa silage, 4% soyhulls, 2.4% mineral-vitamin supplement, and 30% neutral detergent fiber. Supplemental RPM (Mepron, Evonik Corp., Kennesaw, GA) was added to all diets to maintain a Lys:Met ratio of 3.1 in digested AA. Ground high-moisture corn was reduced and soybean meal (SBM) plus RPM increased to give diets containing, on average, 11% CP (28% corn, 31% starch, 6% SBM, 4 g of RPM/d), 13% CP (23% corn, 29% starch, 10% SBM, 8 g of RPM/d), 15% CP (19% corn, 26% starch, 15% SBM, 10 g of RPM/d), and 17% CP (14% corn, 24% starch, 19% SBM, 12 g of RPM/d). Data from the last 14 d of each period were analyzed using the mixed procedures in SAS (SAS Institute Inc., Cary, NC). With the exception of milk fat and milk lactose content, we found no significant effects of diet on all production traits. We did note linear responses to dietary CP concentration for intake, production of milk and milk components, and MUN. Cows fed the 11% CP diet had reduced DM intake, lost weight, and yielded less milk and milk components. Mean separation indicated that only true protein yield was lower on 13% CP than on 17% dietary CP, but not different between 15 and 17% CP. This indicated no improvement in production of milk and milk components above 15% CP. Quadratic trends for yield of milk, energy-corrected milk, and true protein suggested that a dietary CP concentration greater than 15% may be necessary to maximize production or, alternately, that a plateau was reached and no further CP was required. Although diet influenced apparent digestibility of DM, organic matter, and neutral detergent fiber, digestibility did not increase linearly with dietary CP. However, we observed linear and quadratic effects of dietary CP on acid detergent fiber digestibility. As expected, we found a linear effect of dietary CP on apparent N digestibility and on fecal and urinary N excretion, but no effect of diet on estimated true N digestibility. Ruminal concentrations of ammonia, total AA, peptides, and branched-chain volatile fatty acids also increased linearly with dietary CP. Quadratic responses indicated that 14.0 to 14.8% CP was necessary to optimize digestion and energy utilization. Overall results indicated that, when RPM was added to increase Lys:Met to 3.1, 15% CP was adequate for lactating dairy cows fed corn silage diets supplemented with SBM and secreting about 40 kg of milk/d; N excretion was lower than at 17% CP but with no reduction in yield of milk and milk components.

Effects of replacing soybean meal with canola meal or treated canola meal on ruminal digestion, omasal nutrient flow, and performance in lactating dairy cows.

Extrusion treated canola meal (TCM) was produced in an attempt to increase the rumen-undegraded protein fraction of canola meal (CM). The objective of this study was to evaluate the effects of replacing soybean meal (SBM) with CM or TCM on ruminal digestion, omasal nutrient flow, and performance in lactating dairy cows. To assess performance, 30 multiparous Holstein cows averaging (mean ± SD) 119 ± 23 d in milk and 44 ± 7 kg of milk/d and 15 primiparous cows averaging 121 ± 19 d in milk and 34 ± 6 kg of milk/d were blocked in a randomized complete block design with a 2-wk covariate period and 12-wk experimental period (experiment 1). Dietary ingredients differed only in protein supplements, which were SBM, CM, or TCM. All diets were formulated to contain (dry matter basis) 30% alfalfa silage, 30% corn silage, 4% soy hulls, 2.4% mineral-vitamin premix, and 16% CP. The SBM diet contained 25% high-moisture shelled corn and 8.6% SBM; the canola diets contained 22% high-moisture shelled corn and either 11.2% CM or 11.4% TCM. To assess ruminal digestion and omasal nutrient flow, 6 rumen-cannulated cows were blocked into 2 squares of 3 cows and randomly assigned within blocks to the same 3 dietary treatments as in experiment 1 in a replicated 3 × 3 Latin square design (experiment 2). Data were analyzed using the MIXED procedure of SAS (SAS Institute, Cary, NC). Orthogonal contrasts were used to compare effects of different protein supplements: SBM versus CM + TCM and CM versus TCM. In experiment 1, compared with SBM, apparent total-tract digestibilities of dry matter and nutrients were greater in cows fed both CM diets, and there was a tendency for nutrient digestibilities to be higher in cows fed CM compared with TCM. Diets did not affect milk yield and milk components; however, both canola diets decreased urinary urea N (% of total urinary N), fecal N (% of total N intake), and milk urea N concentration. In experiment 2, compared with SBM, both canola diets increased N intake and tended to increase rumen-degraded protein supply (kg/d) and N truly digested in the rumen (kg/d). Diets did not affect ruminal digestibility, efficiency of microbial protein synthesis, and rumen-undegraded protein flow among diets. Results from this experiment indicate that replacing SBM with CM or TCM in diets of lactating cows improved digestibility and may reduce environmental impact. Moreover, under the conditions of the present study, treating CM by extrusion did not improve CM utilization.

Effects of carbohydrate and nitrogen supplementation on fermentation of cheatgrass () in a dual-flow continuous culture system.

Cheatgrass (CG; ), an introduced winter annual grass, is an aggressive invader of the sagebrush community in the Western United States. Because of its greater flammability, mature CG constitutes a fire hazard leading to repeated wildfires. One fuel-reduction strategy is livestock grazing. The objective of this study was to evaluate the effects of urea, molasses, or a combination of urea and molasses supplementation of a CG-based diet on digestibility, microbial fermentation, bacterial protein synthesis, and nutrient flow using a dual-flow continuous culture system. Eight fermenters were used in a replicate 4 × 4 Latin square design with four 10-d experimental periods. Experimental treatments (DM basis) were 1) forage only (CON), 2) CG plus urea alone (URE; 1.36% urea), 3) CG plus molasses alone (MOL; 15.9% molasses), and 4) CG plus urea and molasses combined (URE+MOL; 1.28% urea plus 19.3% molasses). Each fermenter was fed 72 g/d of DM, and data were analyzed using the GLIMMIX procedure of SAS (SAS Inst. Inc., Cary, NC). The true digestibilities of NDF and ADF were not affected by diets ( > 0.05). Molasses-containing diets had greater true digestibility of OM ( = 0.02). However, true digestibility of CP was increased when molasses was fed alone ( < 0.01). Molasses-containing diets had lower pH ( < 0.01) and greater VFA concentrations ( < 0.01) compared to those of the other diets. The URE+MOL diet resulted in a greater VFA concentration ( < 0.01). Propionate concentration increased ( < 0.01), whereas acetate concentration decreased ( < 0.01) when molasses alone or in combination with urea was added to the diets. Supplying molasses alone resulted in greater ( = 0.03) total branched-chain VFA compared to the other diets. The concentration of NH-N and total N flow increased ( < 0.01) in response to urea supplementation and was greater ( < 0.01) when urea alone was supplemented in the diet. On the other hand, molasses-supplemented diets yielded more non-ammonia N ( < 0.01) and bacterial N ( = 0.04). Supplementation had no effect ( = 0.83) on bacterial efficiency. Results from this study indicate that the addition of urea and molasses in a CG-based diet could improve nutrient supply to animals, notably VFA supply and microbial N supply; however, in the levels tested in this study, it did not improve CG utilization as assessed by NDF digestion.

Effects of flaxseed and chia seed on ruminal fermentation, nutrient digestibility, and long-chain fatty acid flow in a dual-flow continuous culture system.

Flaxseed (FS) and chia seed (CS) are oilseeds rich in omega-3 fatty acids, which may change meat and milk composition when added to ruminants' diets and may have health benefits for humans. Literature on the effects of CS supplementation on ruminal metabolism is nonexistent. A dual-flow continuous culture fermenter system consisting of 6 fermenters was used to assess the effect of FS and CS supplementation in an alfalfa hay-based diet on ruminal fermentation, nutrient digestibility, microbial protein synthesis, and long-chain fatty acid flow. Diets were randomly assigned to fermenters in a replicated 3 × 3 Latin square design, with 3 consecutive periods of 10 d each, consisting of 7 d for diet adaptation and 3 d for sample collection. Each fermenter was fed a total of 72 g of DM/d divided in 6 equal portions. Treatments were 1) alfalfa hay + calcium soaps of palm oil fatty acid (MEG; 69.3 g DM/d of alfalfa hay plus 2.7 g DM/d of calcium soaps of palm oil fatty acid), 2) alfalfa hay + FS (FLAX; 68.4 g DM/d of alfalfa hay plus 3.6 g DM/d of ground FS), and 3) alfalfa hay + CS (CHIA; 68.04 g DM/d of alfalfa hay plus 3.96 g DM/d of ground CS). Dietary treatments had similar amounts of total fat, and fat supplements were ground to 2-mm diameter. Effluents from the last 3 d of incubation were composited for analyses. Data were analyzed using the MIXED procedure of SAS. Ruminal apparent and true nutrient digestibility of all nutrients did not differ ( > 0.05) among treatments. Compared with MEG, FLAX and CHIA increased the flows of C18:3 -3, C20:4 -6, and total PUFA ( < 0.01). Both CHIA and FLAX treatments had greater ruminal concentrations of C18:0, indicating that both CS and FS fatty acids were extensively biohydrogenated in the rumen. The NH-N concentration, microbial N flow, and efficiency of microbial protein synthesis were not affected ( > 0.05) by treatments. Lastly, there were no differences ( > 0.05) among diets for total VFA concentration and molar proportions of individual VFA. Results from this study indicate that FS and CS supplementation did not impair ruminal fermentation, digestibility, microbial efficiency, and ruminal N metabolism. Overall, CS appears to be as effective as FS as a fat source when added to ruminants' diets using a dual-flow continuous culture system.

Effects of feeding lauric acid or coconut oil on ruminal protozoa numbers, fermentation pattern, digestion, omasal nutrient flow, and milk production in dairy cows.

The objectives of this study were to evaluate the feeding of coconut oil (CO), in which lauric acid (La) comprises about 50% of the fatty acid composition, as a practical rumen protozoa (RP) suppressing agent, to assess whether the source of La affects ruminal fermentation and animal performance and to test whether suppressing RP improves N utilization, nutrient digestion, nutrient flow at the omasal canal, and milk production. Fifteen multiparous Holstein cows (3 fitted with ruminal cannulas) and 15 primiparous Holstein cows (3 fitted with ruminal cannulas) were used in a replicated 3×3 Latin square experiment with 14d of adaptation and 14d of sample collection. Diets were fed as total mixed ration and contained (dry matter basis) 10% corn silage, 50% alfalfa silage, and 40% concentrate. The control diet contained 3% (dry matter basis) calcium soaps of palm oil fatty acids (Megalac, Church & Dwight Co. Inc., Princeton, NJ) as a ruminally inert fat source and had no added La or CO. Diets with La and CO were formulated to contain equal amounts of La (1.3%, dry matter basis). Dry matter intake was not affected by treatment. Both CO and La reduced RP numbers by about 40%. Lauric acid reduced yield of milk and milk components; however, CO did not affect yield of milk and yields of milk components. Both La and CO caused small reductions in total VFA concentration; CO increased molar proportion of ruminal propionate, reduced ruminal ammonia and branched-chain volatile fatty acids, suggesting reduced protein degradation, and reduced milk urea N and blood urea N concentrations, suggesting improved protein efficiency. Lauric acid reduced total-tract apparent digestibility of neutral detergent fiber and acid detergent fiber as well as ruminal apparent digestibility of neutral detergent fiber and acid detergent fiber as measured at the omasal canal; however, CO did not alter fiber digestion. Microbial protein flow at the omasal canal, as well as the flow of N fractions at the omasal canal, did not differ among treatments. Results from this experiment have confirmed that dietary La is not a practical agent for suppressing RP population in dairy cows, mainly because of its negative effects on fiber digestion and ruminal fermentation. Intake of CO appeared to reduce ruminal and improve protein efficiency, but did not improve milk production, milk composition, or increase microbial outflow from the rumen. Based on the results of this study, a 40% reduction of RP population is not sufficient to improve N utilization in dairy cows.